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Identification and Excretion of Estrogen in Urine during the Estrous Cycle of the Ewe¹

Purdue University, Lafayette, Indiana

Abstract

Estrone was identified in urine of the nonpregnant ewe by comparing its behavior to crystalline standard estrone and by its failure to separate from added ¹⁴C-estrone through solvent partitioning, paper chromatography, successive thin-layer chromatography in different systems, gas-chromatography (GLC) on three columns, derivative formation and chromatography of the derivatives. An infrared spectrum of the trimethylsilyl ether compared favorably with the spectrum of standard estrone trimethylsilyl ether. An estradiol was tentatively identified by several of the above methods but not enough compound was present to obtain an infrared spectrum. The highest average rate of excretion of estrone and estradiol occurred during estrus averaging 394 and 479 µg./24 hr. as compared to the lowest rates of 20 and 17 µg./24 hr., respectively, the second day after estrus. Levels of excretion were intermediate for days 3 through 8 and 13 through 16. Estrone excretion averaged 191 µg./24 hr. for days 9 to 10 compared to 54 on days 7 to 8 and 90 on days 13 to 14. For the 46 samples of urine analyzed from 37 ewes estrone represented 61% of the total estrogen measured. However, estrone represented 45% of the total during estrus, 30% for days 5 to 6, and over 80% from days 13 through 16. Expressing excretion rate as ng./mg. of urinary creatinine was highly correlated (r=0.97–0.98) to total excretion based on total collection of urine for 24 hours.

¹ Journal Paper No. 3730, Purdue University Agricultural Experiment Station. A contribution from the Department of Animal Sciences, 47907. This work was partially supported by Grant No. 3875 from the Purdue Research Foundation, Lafayette, Indiana.

² Present address: Marshfield Clinic Foundation, Marshfield, Wisconsin 54449.